IN VITRO ANTIOXIDANT RESPONSE OF THE EQUINE BLOOD TREATED BY EXTRACT DERIVED FROM LEAVES OF FICUS DELTOIDEA JACK (MORACEAE):

H. Tkachenko; N. Kurhaluk; O. Hasiuk; S. Beschasnyi; L. Buyun; V. Honcharenko; A. Prokopiv

doi:10.32999/ksu2524-0838/2022-33-7

H. Tkachenko
N. Kurhaluk
O. Hasiuk
S. Beschasnyi
L. Buyun
V. Honcharenko
A. Prokopiv

DOI: https://doi.org/10.32999/ksu2524-0838/2022-33-7

Keywords: Ficus deltoidea Jack, lipid peroxidation, oxidatively modified proteins, superoxide dismutase, catalase, glutathione peroxidase, ceruloplasmin, total antioxidant capacity

Abstract

Ficus deltoidea Jack, which has strong antioxidant properties, is widely consumed in traditional medicine as a treatment for various diseases. The current study aimed to investigate the oxidative stress biomarkers, such as 2-thiobarbituric acid reactive substances (TBARS), aldehydic and ketonic derivatives of oxidatively modified proteins, and total antioxidant capacity, as well as antioxidant defenses (activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), ceruloplasmin (CP)) in the equine erythrocytes and plasma to evaluate the antioxidant activities of the aqueous extract derived from leaves of Ficus deltoidea. The leaves of F. deltoidea were collected at M.M. Gryshko National Botanic Garden (Kyiv, Ukraine) and the Botanic Garden of Ivan Franko National University in Lviv (Lviv, Ukraine). Freshly collected leaves were washed, weighed, crushed, and homogenized in 0.1M phosphate buffer (pH 7.4) (in the proportion of 1:19, w/w) at room temperature. The extracts were then filtered and used for analysis. A volume of 0.1 mL of the plant extract was added to 1.9 mL of clean equine erythrocytes or plasma (the final concentration of the extract was 5 mg/mL). For positive control, 0.1 mL of phosphate buffer (pH 7.4) was used. The treatment of equine plasma and erythrocytes by extract derived from leaves of F. deltoidea resulted in reduced carbonyl derivatives of the oxidatively modified protein. The levels of lipid peroxidation were non-significantly changed. The incubation of equine plasma with an extract derived from leaves of F. deltoidea resulted in an increase in the activity of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase with a simultaneous decrease of ceruloplasmin level. The level of total antioxidant capacity was non-significantly increased. The results obtained suggested the antioxidant activity of the extract derived from the leaves of F. deltoidea plants exhibiting a decrease in the biomarkers of oxidative stress and enhancement of antioxidant defenses in the equine erythrocytes and plasma after in vitro treatment with the extract. However, further detailed investigation, especially in vivo and in vitro antioxidant studies is needed to justify the use of extract derived from leaves of F. deltoidea as a natural source of antioxidants.

References

Abrahim NN, Abdul-Rahman PS, Aminudin N. The antioxidant activities, cytotoxic properties, and identification of water-soluble compounds of Ficus deltoidea leaves. PeerJ. 2018;6:e5694. doi: 10.7717/peerj.5694.
Adam Z, Khamis S, Ismail A, Hamid M. Ficus deltoidea: A Potential Alternative Medicine for Diabetes Mellitus. Evid Based Complement Alternat Med. 2012;2012:632763. doi: 10.1155/2012/632763.
Aris SRS, Mustafa S, Ahmat N, Jaafar FM, Ahmad R. Phenolic content and antioxidant activity of fruits of Ficus deltoidea var angustifolia sp. Malaysian J Anal Sci. 2009;13:146-150.
Ashraf K, Haque MR, Amir M, Ahmad N, Ahmad W, Sultan S, Ali Shah SA, Mahmoud Alafeefy A, Mujeeb M, Bin Shafie MF. An Overview of Phytochemical and Biological Activities: Ficus deltoidea Jack and Other Ficus spp. J Pharm Bioallied Sci. 2021;13(1):11-25. doi: 10.4103/jpbs.JPBS_232_19.
Berg CC, Corner EJH. Moraceae (Ficus). In: Noteboom H.P. (ed.) Flora Malesiana, Ser. 1, Vol. 17, Part 2. National Herbarium Nederland, Leiden, 2005, p. 1-730.
Bunawan H, Amin NM, Bunawan SN, Baharum SN, Mohd Noor N. Ficus deltoidea Jack: A Review on Its Phytochemical and Pharmacological Importance. Evid Based Complement Alternat Med. 2014;2014:902734. doi: 10.1155/2014/902734.
Carillon J, Rouanet JM, Cristol JP, Brion R. Superoxide dismutase administration, a potential therapy against oxidative stress related diseases: several routes of supplementation and proposal of an original mechanism of action. Pharm Res. 2013;30(11):2718-2728. doi: 10.1007/s11095-013-1113-5.
Che Ahmad Tantowi NA, Hussin P, Lau SF, Mohamed S. Mistletoe fig (Ficus deltoidea Jack) leaf extract prevented postmenopausal osteoarthritis by attenuating inflammation and cartilage degradation in rat model. Menopause. 2017;24(9):1071-1080. doi: 10.1097/GME.0000000000000882.
Cook JM, Rasplus J-Y. Mutualists with attitude: coevolving fig wasps and figs. Trends in Ecology & Evolution 2003;18(5):241-248.
Dubinina EE, Burmistrov SO, Khodov DA, Porotov IG. Okislitel'naia modifikatsiia belkov syvorotki krovi cheloveka, metod ee opredeleniia [Oxidative modification of human serum proteins. A method of determining it]. Vopr Med Khim. 1995;41(1):24-26. (In Russian).
Galaktionova LP, Molchanov AV, El'chaninova SA, Varshavskiĭ BIa. Sostoianie perekisnogo okisleniia u bol'nykh s iazvennoĭ bolezn'iu zheludka i dvenadtsatiperstnoĭ kishki [Lipid peroxidation in patients with gastric and duodenal peptic ulcers]. Klin Lab Diagn. 1998;(6):10-14 (In Russian).
Garcia YJ, Rodríguez-Malaver AJ, Peñaloza N. Lipid peroxidation measurement by thiobarbituric acid assay in rat cerebellar slices. J Neurosci Methods. 2005;144(1):127-135. doi: 10.1016/j.jneumeth.2004.10.018.
Hakiman M, Maziah M. Non enzymatic and enzymatic antioxidant activities in aqueous extract of different Ficus deltoidea accessions. J Med Plant Res. 2009;3:120-131.
Healy J, Tipton K. Ceruloplasmin and what it might do. J Neural Transm (Vienna). 2007;114(6):777-781. doi: 10.1007/s00702-007-0687-7.
Kamyshnikov VS. A reference book on the clinic and biochemical researches and laboratory diagnostics. MEDpress-inform, Moscow, 2004.
Kelly FJ, Mudway IS. Protein oxidation at the air-lung interface. Amino Acids. 2003;25(3-4):375-396. doi: 10.1007/s00726-003-0024-x.
Koroliuk MA, Ivanova LI, Maĭorova IG, Tokarev VE. Metod opredeleniia aktivnosti katalazy [A method of determining catalase activity]. Lab Delo. 1988;(1):16-19 (In Russian).
Kostiuk VA, Potapovich AI, Kovaleva ZhV. Prostoĭ i chuvstvitel'nyĭ metod opredeleniia aktivnosti superoksid- dismutazy, osnovannyĭ na reaktsii okisleniia kvertsetina [A simple and sensitive method of determination of superoxide dismutase activity based on the reaction of quercetin oxidation]. Vopr Med Khim. 1990;36(2):88-91. (In Russian).
Lansky EP, Paavilainen HM. Figs: the genus Ficus. In: Hardman R. (ed.) Traditional herbal medicines for modern times, Vol. 9. CRC Press, Boca Raton, 2011, p. 1-357.
Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER. Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol. 1990;186:464-478. doi: 10.1016/0076-6879(90)86141-h.
Manurung H, Kustiawan W, Kusuma IW, Marjenah Total flavonoid content and antioxidant activity in leaves and stems extract of cultivated and wild tabat barito (Ficus deltoidea Jack) AIP Conf Proc. 2017;1813:020007.
Martins Alves AM, Pereira Menezes Reis S, Peres Gramacho K, Micheli F. The glutathione peroxidase family of Theobroma cacao: Involvement in the oxidative stress during witches' broom disease. Int J Biol Macromol. 2020;164:3698-3708. doi: 10.1016/j.ijbiomac.2020.08.222.
Misbah H, Aziz AA, Aminudin N. Antidiabetic and antioxidant properties of Ficus deltoidea fruit extracts and fractions. BMC Complement Altern Med. 2013;13:118. doi: 10.1186/1472-6882-13-118.
Mohd Ariff A, Abu Bakar NA, Abd Muid S, Omar E, Ismail NH, Ali AM, Mohd Kasim NA, Mohd Nawawi H. Ficus deltoidea suppresses endothelial activation, inflammation, monocytes adhesion and oxidative stress via NF-κB and eNOS pathways in stimulated human coronary artery endothelial cells. BMC Complement Med Ther. 2020;20(1):56. doi: 10.1186/s12906-020-2844-6.
Mohd Dom NS, Yahaya N, Adam Z, Nik Abd Rahman NMA, Hamid M. Antiglycation and Antioxidant Properties of Ficus deltoidea Varieties. Evid Based Complement Alternat Med. 2020;2020:6374632. doi: 10.1155/2020/6374632.
Moin VM. Prostoĭ i spetsificheskiĭ metod opredeleniia aktivnosti glutationperoksidazy v éritrotsitakh [A simple and specific method for determining glutathione peroxidase activity in erythrocytes]. Lab Delo. 1986;(12):724-727. (In Russian).
Nurdiana S, Goh YM, Ahmad H, Dom SM, Syimal'ain Azmi N, Noor Mohamad Zin NS, Ebrahimi M. Changes in pancreatic histology, insulin secretion and oxidative status in diabetic rats following treatment with Ficus deltoidea and vitexin. BMC Complement Altern Med. 2017;17(1):290. doi: 10.1186/s12906-017-1762-8.
Omar MH, Mullen W, Crozier A. Identification of proanthocyanidin dimers and trimers, flavone C-Glycosides, and antioxidants in Ficus deltoidea, a malaysian herbal tea. J Agric Food Chem. 2011;59(4):1363-1369. doi: 10.1021/jf1032729.
Ooi TC, Ibrahim FW, Ahmad S, Chan KM, Leong LM, Mohammad N, Siew EL, Rajab NF. Antimutagenic, Cytoprotective and Antioxidant Properties of Ficus deltoidea Aqueous Extract In Vitro. Molecules. 2021;26(11):3287. doi: 10.3390/molecules26113287.
Ravin HA. An improved colorimetric enzymatic assay of ceruloplasmin. J Lab Clin Med. 1961;58:161-168.
Rosa AC, Corsi D, Cavi N, Bruni N, Dosio F. Superoxide Dismutase Administration: A Review of Proposed Human Uses. Molecules. 2021;26(7):1844. doi: 10.3390/molecules26071844.
Samsulrizal N, Goh YM, Ahmad H, Md Dom S, Azmi NS, NoorMohamad Zin NS, Ebrahimi M. Ficus deltoidea promotes bone formation in streptozotocin-induced diabetic rats. Pharm Biol. 2021;59(1):66-73. doi: 10.1080/13880209.2020.1865411.
Tkachenko H, Buyun L, Kurhaluk N, Honcharenko V, Prokopiv A, Osadowski Z. In Vitro Antioxidant Activities of Aqueous Extracts Derived from Leaves of Juvenile and Mature Shoots of Ficus pumila L. (Moraceae). Agrobiodivers Improv Nutr Health Life Qual, 2019;(3), 1–13. doi:10.15414/agrobiodiversity.2019.2585-8246.001-013.
Tkachenko H, Buyun L, Osadowski Z, Honcharenko V, Prokopiv A. Oxidative stress biomarkers in the equine plasma and erythrocytes treated in vitro by leaf extract obtained from Ficus religiosa L. (Moraceae). Agrobiodivers Improv Nutr Health Life Qual, 2018;(2), 184-200. doi:10.15414/agrobiodiversity.2018.2585-8246.184-200.
Wiggins JE, Goyal M, Wharram BL, Wiggins RC. Antioxidant ceruloplasmin is expressed by glomerular parietal epithelial cells and secreted into urine in association with glomerular aging and high-calorie diet. J Am Soc Nephrol. 2006;17(5):1382-1387. doi: 10.1681/ASN.2005111239.
Zaid SSM, Othman S, Kassim NM. Protective role of Ficus deltoidea against BPA-induced impairments of the follicular development, estrous cycle, gonadotropin and sex steroid hormones level of prepubertal rats. J Ovarian Res. 2018;11(1):99. doi: 10.1186/s13048-018-0466-0.
Zar JH. Biostatistical Analysis. 4th ed., Prentice-Hall Inc., Englewood Cliffs, New Jersey,1999.
Zolkiffly SZI, Stanslas J, Abdul Hamid H, Mehat MZ. Ficus deltoidea: Potential inhibitor of pro-inflammatory mediators in lipopolysaccharide-induced activation of microglial cells. J Ethnopharmacol. 2021;279:114309. doi: 10.1016/j.jep.2021.114309.